English plantain and psyllium: lack of cross-allergenicity by crossed immunoelectrophoresis

BACKGROUND: English plantain (Plantago lanceolata) weed pollen and psyllium (Plantago ovata) husk dust are inhalant allergens. Because of the phylogenetic relationship between these plant species, cross-allergenicity has been a concern. OBJECTIVE: The purpose of this study was to investigate the possible cross-allergenicity of plantain and psyllium. METHODS: Homologous and heterologous crossed immunoelectrophoresis (CIE) were performed using a commercial English plantain pollen extract and an extract of psyllium seed embryo. Crossed radioimmunoelectrophoresis (CRIE) was performed using sera from subjects who were RAST positive only to plantain (group A), RAST positive only to psyllium (group B), RAST positive to both plantain and psyllium (group C), or RAST negative to both (group D). RESULTS: All of the group A plantain subjects showed IgE binding to at least one of the six plantain allergens in homologous plantain CRIEs while only one of the sera from the group B subjects reacted very weakly to these plantain allergens. In homologous psyllium CRIE, all group B subjects showed pronounced IgE binding to 2 to 7 of the seven psyllium allergens. Several of the plantain subjects demonstrated only very weak binding to psyllium allergens. Heterologous CRIEs demonstrated little relevant IgE binding. CONCLUSIONS: These results indicate that there is little cross-allergenicity between psyllium husk and English plantain pollen.     

Polymorphisms of complement component I and C1R subcomponent of C1 in nine aboriginal Taiwanese populations

Complement component I (IF) and C1R subcomponent of C1 (C1R) types were determined by isoelectric focusing and subsequent immunoblotting techniques for 658 individuals from nine aboriginal Taiwanese populations. The frequency of the IF*A allele ranges from 0.075 (Bunun) to 0.430 (Saisiat), and a new variant allele IF*B2 was found to have polymorphic frequency in the Atayal. The frequency of the C1R*1 allele ranges from 0.410 (Yami) to 0.650 (Atayal), and the frequency of the C1R*2 allele ranges from 0.265 (Atayal) to 0.586 (Saisiat). The C1R*5 allele was found in five populations (Atayal, Bunun, Ami, Puyuma, Yami), and the C1R*9 allele was found in two populations (Tsou, Puyuma). The results indicate a remarkable degree of genetic variability among these populations. The variability may reflect long-term genetic and geographic isolation of each population.     

X-Prolyl dipeptidyl-aminopeptidase activity, with X-proline p-nitroanilides as substrates, in normal and pathological human sera

X-Prolyl dipeptidyl-aminopeptidase (no EC no. assigned) activity in normal and pathological human sera was assayed with several newly synthesized X-proline p-nitroanilides as chromogenic substrates. Normal values for 88 healthy subjects (15 to 81 years old), with glycylproline p-nitroanilide as substrate at pH 8.7, were 54.9 +/- 1.5 (SE) (range, 25.7 - 96.0) mumol/min per liter of serum at 37 degrees C. The results suggest that the enzyme activities with all X-proline p-nitroanilides were increased in patients with hepatitis and decreased in patients with gastric cancer. On Sephadex G-200 column chromatography, normal human sera showed a single peak of enzyme activity with glycylproline p-nitroanilide as the substrate, which coincided with the peak with glycylproline beta-naphthylamide but was different from the peaks with leucine beta-naphthylamide. Sera from patients with hepatitis or liver cirrhosis showed an increase in the normal peak and the appearance of another new peak with glycylproline p-nitroanilide as substrate.     

Semiautomated microtiter plate assay for monitoring peptidylprolyl cis/trans isomerase activity in normal and pathological human sera

Elements mediating VanA glycopeptide resistance in 106 diverse enterococci from humans and nonhuman sources were compared with the prototype VanA transposon, Tn1546, in Enterococcus faecium BM4147. The isolates included 64 from individual patients at 15 hospitals in the United Kingdom (isolated between 1987 and 1996) and 42 from nonhuman sources in the United Kingdom (27 from raw meat, 7 from animal feces, and 8 from sewage). VanA elements were assigned to 24 groups (designated groups A to X) with primers that amplified 10 overlapping fragments of Tn1546. Ten groups of elements were found only in human enterococci, eight groups of elements were unique to nonhuman strains, and six groups of elements were common in enterococci from all sources. Elements indistinguishable from Tn1546 (group A) were observed more frequently in enterococci from nonhuman sources (34 versus 9%) but were identified in enterococci that caused outbreaks in hospital patients between 1987 and 1995. The most common group found in human enterococci (group H; 33%) was rarely observed in enterococci from other sources (5%). Group H elements differed from Tn1546 in three regions and included a novel insertion sequence, designated IS1542, between orf2 and vanR. The VanA elements of 14 other groups had a similar insertion at this position and/or distinct insertions at other positions. We conclude that VanA elements in enterococci are heterogeneous, although all show regions of homology with Tn1546. Furthermore, the elements most common among the human and nonhuman enterococci studied were different. This approach may be useful for monitoring the evolution of VanA resistance and may also be applicable in local "snapshot" epidemiological studies. However, as transposition events involving insertion sequences accounted for the differences observed between several groups, the stability of the elements must be assessed before their true epidemiological significance can be determined.     

Variability of impedivity in normal and pathological breast tissue

Bite wounds have a special position in traumatology because of their high complication rate compared to similar soft tissue wounds caused by other reasons. The authors report in a retrospective study about the results of 525 patients with bite wounds. In 98 (18.7%) cases the wounds were sutured primarily after surgical revision when there was no sign of infection. Antibiotic therapy (a combination of amoxicillin and clavulan acid) was given to 109 patients (20.8%) who had infected wounds or who were at greater risk for infection (e.g., extremely large wounds, large hematoma). Antibiograms were only made when the wounds had already been infected. The total infection rate came to 11.8%, with 5.2% infected facial bites, 11.3% lower extremity, 18.6% upper extremity and 18.8% hand bite wounds. The infection rate after primary suture was 10.2% (3.2% at the face, 18.8% at the upper extremity and 25% each in the hand and the lower extremity). The average time period from the trauma to the first medical treatment amounted to 11 h in the infected wounds and 2 h in the non-infected ones. Cat bites became infected in 37.5%, dog bites in 14.9%.     

Autoantibodies to M2 mitochondrial autoantigens in normal human sera by immunofluorescence and novel assays

Primary biliary cirrhosis (PBC) is characterized by the presence of antimitochondrial antibodies (anti-M2), directed against the E2 subunits of the 2-oxo-acid dehydrogenase complexes (2-OADC), chiefly pyruvate dehydrogenase complex (PDC-E2). We present here a detailed study, based on a large panel of normal sera, of the specificity of tests for anti-M2 by immunofluorescence and for anti-PDC by other assays for the diagnosis of PBC. The assays for anti-PDC included immunoblotting with bovine heart mitochondria, ELISA using recombinant PDC-E2 and an enzyme inhibition assay using purified porcine PDC. The positivity rates for normal sera were 0 (0/170), 2 (4/201), 1.5 (3/198) and 0% (0/186) for immunofluorescence, immunoblotting, ELISA and the enzyme inhibition assay, respectively. The seven positive reactions detected either by immunoblotting (n = 4) or ELISA (n = 3) were negative by the other three assays and in no instance did biochemical indices give any indication of chronic liver disease. Thus, as judged by reactivity with normal sera, the specificity of a positive test for the antibody to the major M2 autoantigen (PDC-E2) is 100% for immunofluorescence and the enzyme inhibition assay, 98% for immunoblotting and 98.5% for ELISA.     

Langerhans cells in normal and pathological vocal cord mucosa

The Langerhans cells in samples of histologically normal and pathological vocal cord mucosa were counted after identification using S-100 polyclonal antibody. Langerhans cells were commonly seen in vocal cord polyp epithelium but were infrequent in normal cord mucosa. They were also identified in samples of squamous carcinoma, severe dysplasia and chronic inflammation.     

Dynamic structure of subtilisin-eglin c complex studied by normal mode analysis

Normal mode analysis of subtilisin-eglin c complex was performed to investigate the dynamics at the interface between the enzyme and the inhibitor. The internal motions of the complex calculated from the normal modes were divided into three parts: the internal motions changing the shape of each molecule, the external rigid-body motions changing their mutual dispositions, and the coupling between the internal and external motions. From the results of the analysis, the following characteristic features were found in the dynamics at the interface regions: 1) negative correlation between the internal and external motions within each molecule, and 2) positive correlation between the external motions of the two molecules. The former decreases the apparent amplitudes of motions at the interface. The latter minimizes the interference between individual motions of the two molecules. These dynamic characteristics allow the enzyme and the inhibitor to move as freely as possible. This finding suggests that the experimental evidence of the large entropy gain on binding should be attributed not only to strong hydrophobic interactions, but also to the dynamic structure of the complex, which is found to minimize an unavoidable loss of the conformational entropy on binding.     

The first subcomponent of complement, C1q, triggers the production of IL-8, IL-6, and monocyte chemoattractant peptide-1 by human umbilical vein endothelial cells

We and others have demonstrated previously the occurrence of cC1qR/CaR, a receptor for the collagen-like stalks of complement component C1q, on endothelial cells. In the present study we investigated whether binding of C1q to endothelial cells resulted in enhancement of cytokine or chemokine production. HUVEC produced 82 +/- 91 pg/ml of IL-8, 79 +/- 113 pg/ml of IL-6, and 503 +/- 221 pg/ml of monocyte chemoattractant peptide-1 (MCP-1) under basal conditions. Incubation with C1q resulted in a time- and dose-dependent up-regulation of IL-8 (1012 +/- 43 pg/ml), IL-6 (392 +/- 20 pg/ml), and MCP-1 (2450 +/- 101 pg/ml). This production is dependent on de novo protein synthesis, as demonstrated by the detection of specific mRNA after C1q stimulation, and inhibition of peptide production in the presence of cycloheximide. The production of all factors was inhibited (69 +/- 7%) by the collagenous fragments of C1q, while the C1q globular heads only induced 13 +/- 11% inhibition. When HUVEC were incubated with C1q in the presence of aggregated IgM, enhanced production of IL-8 (2500 +/- 422 pg/ml), IL-6 (997 +/- 21 pg/ml), and MCP-1 (5343 +/- 302 pg/ml) was found. Furthermore, F(ab')2 anti-calreticulin partially inhibited the production of IL-8, confirming at least the involvement of cC1qR/CaR. These experiments suggest that in an inflammatory response C1q not only is able to activate the complement pathway, but when presented in a proper fashion also might induce the production of factors that contribute to acute phase responses and recruitment of inflammatory cells.     

Detection of hydatid antigen in urine by countercurrent immunoelectrophoresis

Hydatid antigen was demonstrated for the first time in the urine of patients with hydatid disease by countercurrent immunoelectrophoresis (CIEP). The antigen was detected in the concentrated urine of 7 of 16 (43.75% positive) patients with surgically confirmed hydatid disease, 4 of 10 (40% positive) patients with ultrasound-proven hydatid disease (daughter cysts or prominent septation and hydatid sands demonstrated by ultrasound), and 8 of 14 (57.14% positive) patients with clinically diagnosed (presumptive) hydatid disease. No antigen was detected in the concentrated urine from 24 patients with parasitic diseases other than hydatid disease. However, antigen was detected in 2 (8% false positive) of 25 concentrated urine samples collected from healthy control subjects (blood donors and students). These result suggest that the detection of hydatid antigen in the urine by CIEP is a simple, rapid, and noninvasive method of diagnosis of hydatid disease.     

Time series analysis of glottal airflow in normal and pathological phonation

Time series analysis of glottal airflow was carried out on 26 normal controls and 40 patients with voice disorders, using a modification of Isshiki's original technique which uses a hot-wire flowmeter, taking cycle-by-cycle fluctuations into consideration. The mean flow rate and mean AC/DC were shown to have significant differences among normal and patient groups. The standard deviations of AC/DC and AC/DC perturbation were calculated from the AC/DC value of 50 cycles and shown not to vary significantly among the normal and patient groups. The relationship between AC/DC and perceptual impression of voice was also studied among 20 selected patients with breathy voices. Using Spearman's rank correlation coefficient, this relationship was found to have statistical significance (P < .05).